Friday, September 28, 2012

MMP9 and height

MMP9 regulates the cellular response to inflammation after skeletal injury.

"To assess the role of inflammation on skeletal cell differentiation, we used mouse models of fracture repair that stimulate either intramembranous or endochondral ossification. In the first model, fractures are rigidly stabilized leading to direct bone formation, while in the second model, fracture instability causes cartilage and bone formation. We compared the inflammatory response in these two mechanical environments and found changes in the expression patterns of inflammatory genes and in the recruitment of inflammatory cells and osteoclasts. We then exploited matrix metalloproteinase 9 (MMP9) that is expressed in inflammatory cells and osteoclasts, which is a potential regulator of cell fate decisions during fracture repair. Mmp9-/- mice heal stabilized fractures via endochondral ossification, while wild type mice heal via intramembranous ossification[So to grow taller being MMP knockout may be better]. In parallel, we observed increases in macrophages and T cells in the callus of Mmp9-/- compared to wild type mice. To assess the link between the profile of inflammatory cells and skeletal cell fate functionally, we transplanted Mmp9-/- mice with wild type bone marrow, to reconstitute a wild type hematopoietic lineage in interaction with the Mmp9-/- stroma and periosteum. Following transplantation, Mmp9-/- mice healed stabilized fractures via intramembranous ossification and exhibited a normal profile of inflammatory cells. Mmp9-/- periosteal grafts healed via intramembranous ossification in wild type hosts, but healed via endochondral ossification in Mmp9-/- hosts. We observed that macrophages accumulated at the periosteal surface in Mmp9-/- mice, suggesting that cell differentiation in the periosteum is influenced by factors such as BMP2 that are produced locally by inflammatory cells. MMP9 mediates indirect effects on skeletal cell differentiation by regulating the inflammatory response and the distribution of inflammatory cells, leading to the local regulation of periosteal cell differentiation."

The mice were 12-16 weeks old. Mast cells(connective tissue cell) typically produce MMP9. Fractures were produced by three point bending loading.

"Inflammatory mediators such as tumor necrosis factor-α (TNFα) are required for bone formation but can also impair later stages of repair by stimulating cartilage degradation"

"The role of Cox2 is counterbalanced by 5-lipoxygenase (5-LO) and leukotriene inhibitors can stimulate cartilage and bone formation in the early phase of repair through direct actions on chondrocytes"<-Grow taller with leukotriene inhibitors? Pharmacological inhibition of 5-lipoxygenase accelerates and enhances fracture-healing. states that 5-LO inhibition via AA-861 increased COL2, COX2, COL10, and osteopontin. Celecoxib[COX2 inhibitor] also increased COL2.

"MMP9−/− but not wild type skeletal progenitors differentiate into chondrocytes in both non-stabilized and stabilized fractures"

"the presence of Mmp9−/− inflammatory cells alone was not sufficient to induce chondrogenic cell differentiation at the fracture site. "

"Osteoclast recruitment was impaired in Mmp9−/− fracture calluses and bone marrow transplants rescued the cartilage remodeling defects in non-stabilized fractures via providing MMP9-expressing osteoclasts. This result was in concordance with the rescue of the growth plate phenotype in Mmp9−/− developing long bones"

"the recruitment of T cells and macrophages was affected [in] Mmp9−/− fracture calluses"

"This indirect effect of MMP9 on periosteal cell fate was via the regulation of macrophage localization. Only macrophages were located within the periosteum to locally influence chondrogenic and osteogenic cell differentiation. CD4 + T cells were found away from the periosteum, and so are unlikely to have an effect on chondrogenesis and osteogenesis."

"MMP9 acts indirectly at the level of the periosteum"

"Ectopic localization of macrophages in the periosteum of Mmp9−/− stabilized fractures may be responsible for the induction of endochondral ossification as we found an increase in BMP2 expression."

"MMP9 indirectly regulates angiogenesis by regulating the bioavailability of VEGF"

"the lack of MMP9 in both the periosteum and inflammatory cells was responsible for the cartilage phenotype."

Genes upregulated in Unstablized versus Stabilized(MMP9 knockout) at Day 7 that were also upregulated(or -down) by LSJL:

All below are signal transduction related molecules:

COL10A1

C1QTNF3

COL12A1

COL11A1

COL6A1

PTN

MOXD1

ASPN

EDIL3

NOV

LRIG3(down)

TNN

COL16A1

EMILIN3

DKK3

COL1A1

PTGS2

MATN4

HTRA1

Col9A1

MATN3

Col9A2

SMOC2

THBS3

CSPG4

THBS2

CCL21C

COL2A1

KERA

Glycoprotein Related:

TNMD

COL13A1

PRSS35

EPHA3

GPR88(down)

Car12

HAPLN1

Cell Differentiation:

Sox9

COL13A1

ALDH1A3(down)[an enzyme produced by retinoic acid]

NR4A2

SLITRK6

OTHER:

WNT2

LSJL was highly homologous to the most chondrogenic stimulation group in the study MMP9-/- unstabilized fractgure.

Genes upregulated in stabilized versus unstabilized MMP9 KO mice that were also altered in LSJL
So these are essentially downregulated by unstabilized mice:

CCL2(up)
CXCL5
CXCL1(up)
IL6(up)
CD14(up)

Unstabilized non-MMP9 knockout on day 7 versus pre-fracture gene upregulation that were also upregulated(or -downregulated) by LSJL:

Saa3

CCL2

CXCL1

IL6

SERPINA3N

CXCL5(down)

CCL7

CD276

CFI

ANXA8

At day 2 these are the genes that are also upregulated by LSJL that were not previously reported above:

IL1RL1

LSJL produces a similar gene expression profile to MMP9-/- and Unstabilized fracture mice which were pro-chondrogenic thus providing further evidence that LSJL is pro-chondrogenic. No change in MMP9 expression was reported due to LSJL.

From what I gather about the lsjl research, it was done on mice whose epiphisis were not yet closed. Lsjl has not been scientifically proven on adults of any species.

It initially seemed like it may work so the blogger claimed that he tried it and that it did work on him, in order to entice others to try it and experiment on themselves as well.

It may yet work but it seems there was an attempt to recruit people into experimenting on themselves and those people needed to believe that it could work in order to put themselves through the experimentation.

There was even a post here called "Can lsjl work on adults?" There seems to be doubts even with the biggest proponent of LSJL.

tyler tell me how i can help you! serously we need to speed up progress here, your doing an amazing job compiling research.. but wheres the working method?!

just tell me how i can help you make things happen faster here, i really want a lsjl method that would give 2-3 inches! or is that a wrong expectation?? but please, inform us if you can. what is the expectations you expect from LSJL?? the other best option left seems to be distraction osteogenesis. why does that one cause such rapid growth?

You can test LIPUS or provide alternative testing to LSJL. The problem is mechanosensativity so I can't test more than one method at a time because I might not get results with one method just because the cells like sensitivity.

3-5 minutes with at least 100N of load(that's the number they put out in their patent for humans). 100N could be ten pound pounds if you push it down with 20 times the force of gravity. I wonder if there's a way we can measure force.

If you have lab connections there are multiple ways you can help.

LSJL should have unlimited growth potential but adaptation to load is the biggest problem.

Growth Response seems to be based on peak load and distraction osteogenesis causes the largest stimulus via fracture.

yeah i think the best way to help this progress faster and successful we need to do LSJL @ ourselves and as mentioned by tyler ADAPTATION to load is biggest problem and tyler i think you should mention the answer to------->Can you explain the difference between static and dynamic pressure? It seems that compressing the bone continuously for a period of time would be static pressure and dynamic pressure involves force applied for short intervals several times per second. --------ALL THE BEST GUYs

Can you explain the difference between static and dynamic pressure? It seems that compressing the bone continuously for a period of time would be static pressure and dynamic pressure involves force applied for short intervals several times per second.

I've asked this question before but didn't get a response. Sorry for the bother!

My point then is that the method of compression using a clamp described in the "experimental LSJL routine" seems to be static pressure. I've read in multiple places though that static compression actually tends to have a negative effect on chondrocyte proliferation. Tapping would seem to be more akin to dynamic pressure.

My point then is that the method of compression using a clamp described in "Experimental LSJL routine" seems to be static pressure. I've read in several papers though that static pressure has a negative effect on chondrocyte proliferation while dynamic pressure generally has a positive effect. Tapping which would seem more akin to dynamic pressure.

How expensive is a LIPUS machine that you could use at your home and that is powerful enough to get the results you are seeking should your experimentation with ultra sound and lsjl work?

About $1500? Can you afford that? I know people that spend more than that on soft drinks and coffee in a year.

If LIPUS can take lsjl to the next level then you should make it your goal to get a lipus machine. Would you need 2 of them? One for each leg? I won't go into it in this post but, if you could reliably lengthen people's legs by an inch or more, money would be the last of your worries. The problem now is that people aren't certain that you actually increased your leg length at all, and it's hard to tell anything definitive in the photos. And, if you did, why can't others...or why so very few others? The other thing is that most people won't want their ankles or knees widened to any deformed looking degree in the process . People don't want to trade one problem for another. Also, symmetrical results would be expected. Younger people on the forum here might not care about deforming their ankles/knees and having one leg end up being shorter than the other but most adults would find that unacceptable.

would it be possible to move the water with like pulsed magnetic fields? or something like a pulsed wave? i know you mention lipus, but we need to cause the water to move right? so we need something that can influence its movement outside of the body, unless there was a way to do it inside,OR maybe just focus on getting stemcells, if that is the point.

LIPUS has been validated on adult stem cells in vitro to induce chondrogenesis. PEMF hasn't, I believe some electrical stimulation has shown validity however.

PEMF is definitely worth testing. I think maybe in conjunction with LSJL as the genes that it upregulates are different but that could be due to different species and ages. Tommorrow I'll go over all the electricity posts.

It seems this post is getting quite a few responses. The best approach seems to be able to induce as much chondrocytes in the epiphysis as possible and hope they can remodel the bones by stretching it out

remember the law is that the more you remodel the bones, the harder and more adaptable the bones become to external stimuli.

my guess it that If we can combine our brains, we should be able to get a working less invasive faster surgical method within 10-20 years. i have no idea if you did grow that much. my coworker doesn't believe you. if you did, then you are doing something right.

The whole idea of lsjl is supposed to be NON-surgical bone growth, not less invasive faster surgical methods. Basically, "weight training" for the bones.

It doesn't necessarily have to be anything close to a 10 to 20 year ordeal. I think the problem with experimenting in this fashion is that 'height increase' attracts a lot of kids, for obvious reasons, but not enough adults. You can see immaturity in a lot of the posts on the forum. Adults are resigned to the "fact" that they're not going to grow any taller. If the word gets out to enough adults that height increase is a possibility then the entire dynamic changes dramatically. There needs to be a combination of (1)grown ups, (2)people who are familiar with the studies and have a scientific background,(3)adults that have reached their full natural height but are interested in taking it further(4)people willing to test things out themselves.

Tyler, keep in mind that you want to bring this to attention of adults. Right now adults aren't interested as much or at all because most have accepted that their current height not going to ever increase. Adults have the money. Adults equal funding. If you have 3000 or so serious adults reading your blog then you could easily raise whatever money you need to conduct research. There are dozens of lift wearing celebrities that would pay 6 figures if you gave them an inch in height. Adults, Tyler, Adults! But you have to reach that audience and as good as a job you're currently doing it could be better and height increase could be your full time job.

Height Increase is an attractive place for research because there's not a lot of research by adults compared to how easy it is say compared to cancer research.

But that also makes it hard to find resources.

If you have an idea of how to reach adults then please share. According to the Analytics data scientists(there's been a couple visits from West Lafayatte which is where Yokota is in Purdue) do visit the site sometimes but they bounce after only a few seconds. They won't give the site a chance. I've tried tweeting celebrities with height complexes like Ryan Seacrest but no luck.

maybe you should seek them instead of waiting for them to stay longer on the site, it would be better to just keep sending that collaboration request/or something of interest to them and proof of your ideas, just badger them until they give in.

best comments ever...i like this blog because i spent a lot of money for height increase scams like yoko height mfIII growtaller growthflexetc etc.AND ALL SCAMS ARE MENTIONED HERE BY TYLER...i know for sure tyler is researcher and well said that its good topic or place to do research as compared to others like carcinomas and common leading causes of deaths esp heart diseases stroke and breast cancerwe should try to involve adults and celebrities too BUT its gona take time and WE HAVE TO BE PATIENT ..its NOT LIKE A MAGIC MEDICINE that we take and gona see results quickly..a lot needed to be done...AND FOR ALL USERS we should contribute positively to this research as a orthopedic i am sure there is a connection b/w LSJL/LIPUS and osteogenesis/chondrogenesis...goood luck with work..

i dont see why the scientist dont want to speak with you tyler? what do you think is the reason for that seriously why would they ignore this site? this site has done mostly everything right to look credible, you show lots of science and your researching "their" lsjl method they created, why would they ignore your questions then? perhaps if the scientist doesnt want to speak then maybe his assistants or lab partners may be more open to discussion? i just dont see why the scientist wouldnt be open to discussing with you, perhaps he cant read english? need a site translation? i just dont understand it, it doesnt seem right.